Crypthecodinium cohnii (Seligo) Javornicky (ATCC® 30556)

Organism: Crypthecodinium cohnii (Seligo) Javornicky  /  Depositor: CA Beam, M Himes

Permits and Restrictions

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Strain Designations To (Tolo)
Application
Biofuel production
Biosafety Level 1
Isolation
shoreline organic debris, Tolo Harbor, Hong Kong, 1976
Product Format frozen
Type Strain no
Comments
Minor Sibling Species To
genetic diversification
sexuality and meiosis
Medium ATCC® Medium 460: A2E6 medium
Growth Conditions
Temperature: 25.0°C
Duration: axenic
Protocol: ATCCNO: 30021 SPEC: This culture is routinely shipped as a frozen stabilate. Thaw the ampule and aseptically transfer the material to 5 ml of ATCC medium 460 in a 16 x 125 mm screw-capped test tube. Do not distribute the thawed material to a larger volume of medium. It is essential to first establish the culture in a small volume. Screw cap on tightly, loosen one half turn, and incubate culture upright at 25C. The culture should be ready to subculture in approximately 7 days. To subculture, screw the cap on tightly, invert the culture 5 times and aseptically transfer a 0.1 ml aliquot to 5 ml of ATCC medium 460 and incubate as above. Prepare two subcultures weekly. Retain all cultures for up to one month to ensure against loss.
Subcultivation
Protocol: ATCCNO: 30021 SPEC: This culture is routinely shipped as a frozen stabilate. Thaw the ampule and aseptically transfer the material to 5 ml of ATCC medium 460 in a 16 x 125 mm screw-capped test tube. Do not distribute the thawed material to a larger volume of medium. It is essential to first establish the culture in a small volume. Screw cap on tightly, loosen one half turn, and incubate culture upright at 25C. The culture should be ready to subculture in approximately 7 days. To subculture, screw the cap on tightly, invert the culture 5 times and aseptically transfer a 0.1 ml aliquot to 5 ml of ATCC medium 460 and incubate as above. Prepare two subcultures weekly. Retain all cultures for up to one month to ensure against loss.
Cryopreservation

1.   Harvest cells from cultures which are at or near peak density.  Aseptically transfer cells to 15 ml plastic centrifuge tubes and centrifuge at ~150 x g for 5 min.

2. Adjust the concentration of cells to 2 x 106/ml with fresh medium, then dilute to half this concentration by adding an equal amount of a 15% (v/v) sterile glycerol solution in fresh  medium.

3.  Dispense in 0.5 ml aliquots into 1.0 - 2.0 ml sterile plastic screw-capped cryules (special plastic vials for cryopreservation).  The time from mixing of the cell suspension and the glycerol solution, before the cooling cycle begins, should be no less than 15 min and no greater than 30 min.

4.   Place vials in a controlled-rate freezing unit. From room temperature, cool at -1°C/min to -40°C. If freezing unit can compensate for the heat of fusion, maintain rate at -1 C/min through heat of fusion. At -40°C plunge ampules into liquid nitrogen. Alternatively, place the vials in a Nalgene 1°C freezing apparatus.  Place the apparatus at -80°C for 1.5 to 2 hours and then plunge ampules into liquid nitrogen.  (The cooling rate in this apparatus is approximately -1°C/min.)   

5.   The frozen preparations should be stored in either the vapor or liquid phase of a nitrogen refrigerator. Frozen preparations stored below -130°C are stabile indefinitely. Those stored at temperatures above -130°C are progressively less stabile as the storage temperature is elevated. Vials can be stored between -80 and -70°C for no longer than one week.

6.   To establish a culture from the frozen state place an ampule in a water bath set at 35°C. Immerse the vial to a level just above the surface of the frozen material. Do not agitate the vial.

7.   Immediately after thawing, do not leave in the water bath; aseptically remove the contents of the ampule and transfer to a fresh tube (or T25 flask) containing 5 ml of ATCC Medium 460. Incubate culture (if in a tube, incubate vertically) at 20-25°C with the cap loosened one-half turn.  Subculture every 10-14d.

Name of Depositor CA Beam, M Himes
Year of Origin 1976
References

Japanese Patent 7,008,221

Salt GW. Changes in the cell volume of Didinium nasutum during population increase. J. Protozool. 22: 112-115, 1975.

Beam CA, Himes M. Sexual isolation and genetic diversification among some strains of Crypthecodinium cohnii-like dinoflagellates evidence of speciation. J. Protozool. 24: 532-539, 1977.

Seafood products having good taste containing marine microalgae, particularly Crythecodinium cohnii containing docosahexaenoic acid. Japanese Patent 7,008,221

Iizuka T, et al. Marine micro-algae food material containing docosahexaenoic acid, food containing the same and manufacturing method therefor. US Patent 5,547,699 dated Aug 20 1996

Beam CA, Himes M. Sexuality and meiosis in dinoflagellates. In: Beam CA, Himes M, Biochemistry and physiology of protozoa, 2nd ed.3 New York Academic Press: 171-206, 1980.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation